Posters

Constitutive TNF-alpha expression is characteristic of the malignant ovarian surface epithelium. Adenoviral mutants hold great promise as gene therapy vectors but their efficacy is hindered by an inflammatory cascade orchestrated by TNF-alpha. We found that delivering TNF-alpha shRNA to ovarian cancer cells using oncolytic adenoviruses could reduce the inflammatory cascade generated by adenoviruses and also had direct anti-tumour activity on the cancer cells.

CF is caused by mutations in the gene encoding for CFTR. CFTR functions as chloride channel on the apical membrane of epithelia thereby regulating the transport of chloride and also sodium ions indirectly. It seems that the regulation of ENaC fails due to the mutated CFTR protein. And it is assumed that ENaC plays a role in the pathogenesis of chronic lung disease in CF-patients.

In this work, a pressurized bioreactor was used for Y. lipolytica batch cultivation under increased air pressure from 1 bar to 6 bar. Cell growth was strongly enhanced by the pressure rise. The increase of oxygen availability caused the induction of the antioxidant enzyme SOD. The pre-growth of Y. lipolytica under increased pressure conditions did not affect the lipase production ability of the cells.

384-well plate “mini-tube” consumables have been developed by automation companies such as REMP and The Automation Partnership (TAP) to hold compound samples divided into single-use storage tubes. Each tube holds 40 -75 µL, and is filled, sealed, stored and then thawed just once before use. However, when accessed by conventional pipetting technology, the tube’s practical working volume is as little as 20 µL, which means high compound wastage. The seal must also be pierced prior to pipetting to p

Nodal involvement in bladder cancer is an independent indicator of prognosis. This study employed an iterative machine learning process called genetic programming on quantitative expression values of 70 genes to classify primary urothelial carcinoma samples into those associated with or without nodal metastasis. The generated rules showed a strong predilection for ICAM1, MAP2K6 and KDR resulting in gene expression motifs that cumulatively suggested a pattern ICAM1>MAP2K6>KDR for node positive ca

How to Perform High Throughput siRNA TransfectionMark Hewitson, Hanno Hermann and Neil Benn

We demonstrate the optimization of siRNA transfection process for time and costs in a high throughput application with two independent measurements of the RNAi effect: The visual monitoring of Eg5 expression and the quantitative PCR measurement of GAPDH mRNA after transfection of Hela cells. The results show, that 1 wash cycle is sufficient for the removal of any remnants of the transfection complex from disposable tips, making the tips re-usable.

We have developed a competitive qPCR method in which siRNA competes with a homologous forward primer to bind template DNA, giving siRNA concentration dependent inhibition. The addition of E6-siRNA to cqPCR led to inhibition of amplification in a linear concentration-dependent manner, with as little as 200pg of siRNA capable of being detected. Irrelevant siRNA had no effect on amplification confirming specificity

We have developed a competitive qPCR method in which siRNA competes with a homologous forward primer to bind template DNA, giving siRNA concentration dependent inhibition. The addition of E6-siRNA to cqPCR led to inhibition of amplification in a linear concentration-dependent manner, with as little as 200pg of siRNA capable of being detected. Irrelevant siRNA had no effect on amplification confirming specificity

The purpose is to study the role of multidrug resistance-associated protein 5, MRP5 in drug metabolism in human retinal pigment epithelium (RPE). RPE forms the outer part of blood-retinal barrier (BRB) which restricts movements of solutes from systemic bloodstream to the neural retina. The efflux protein, MPR5 is expressed in RPE but its functions are mainly unknown. SiRNA will be tested as a tool to clarify the role of MRP5.